Molecular Devices SpectraMax M3 User Manual

Operation
48 0112-0115 F
3. Choose the emission filter:
 Select an emission cutoff filter that blocks as much of the
residual excitation light as possible without unduly reducing
the fluorescence signal. The cutoff wavelength choices are
325, 420, 435, 475, 495, 515, 530, 550, 570, 590, 610,
630, 665, or 695 nm. The cutoff value should be near the
maximum emission wavelength (preferably between the
excitation wavelength and the maximal emission
wavelength) but at least 10 nm less than the emission
wavelength. If you have questions about this procedure
please contact MDC Technical Support and ask to speak to
an applications scientist.
4. Perform emission scan #2:
 In the SoftMax Pro Software, set up a third plate section for
an emission scan as specified in Step 2 above, except
selecting Manual Cutoff Filter and setting the wavelength to
that determined in Step 3.
 Perform the scan and view the results as a plot of
fluorescence vs. emission wavelength. Note the wavelength
giving the maximum emission (the optimal emission
wavelength).
 Compare the spectra of the sample containing the
fluorophore to the spectra of the buffer blank to get an
estimate of the signal-to-noise ratio. If there is significant
background interference, repeat Steps 3 and 4 with another
choice of cutoff filter.
5. Results
The optimal excitation and emission wavelengths are those
determined in Steps 1 and 4, above.
6. Comments
 In endpoint or kinetic fluorescence modes, the “Autofilter”
feature generally selects the same cutoff filter wavelength as
the above optimization method. If desired, however, you
may specify the cutoff filters manually.
 For emission wavelengths less than 325 nanometers,
experimental iteration is usually the best method of
determining the optimal emission and excitation
wavelengths. Begin optimization by performing Steps 1–4
above. Try emission and excitation wavelength combinations
with the 325 cutoff or with no cutoff filter. Similarly, for
excitation wavelengths greater than 660 nanometers, try
emission and excitation wavelength combinations with the
695 cutoff or with no cutoff filter.

Molecular Devices SpectraMax M3 Specifications

General

Brand	Molecular Devices
Model	SpectraMax M3
Category	Laboratory Equipment
Language	English